TW201332953A - Novel arylamide derivatives having antiandrogenic properties - Google Patents

Abstract

The invention relates to novel arylamide derivatives having formula (I) and stereoisomers and pharmaceutically acceptable salts thereof, where R2, R8, R9 and R11 are as defined in the claims. The arylamide derivatives of formula (I) have antiandrogenic properties. The invention also relates to compounds of formula (I) for use as a medicament and to pharmaceutical compositions comprising them and to their preparation.

Description

Novel arylamine derivatives with antiandrogenic properties

The present invention relates to novel arylamine derivatives, their preparation, pharmaceutical compositions comprising them, and their use in the treatment of androgen receptor-related disorders, such as benign prostate hyperplasia and cancer, particularly It is prostate cancer and/or castration-resistant prostate cancer.

Androgen is produced by testis and adrenal glands, and they play a key role in the development and physiology of normal prostate. The pathogenic principle of benign prostatic hyperplasia (BPH) and prostatic neoplasia, which can progress to adenocarcinoma, is androgen-dependent. The treatment of BPH and prostate cancer (PCa) is chosen to reduce androgen action in the prostate. In fact, almost 90% of men between the ages of 40 and 90 will evolve into BPH or PCa. PCa is the second leading cause of cancer-related death in men and the most commonly diagnosed malignancy. PCa is still incurable under metastatic disposal. Because the incidence of prostate cancer increases with age, the number of newly diagnosed cases continues to rise due to the increase in the life expectancy of the population.
The traditional initial treatment for PCa is hormone or androgen deprivation therapy (ADT). The experimental ADT was first described in 1941. The first line of advanced PCa is recognized worldwide by surgical castration or via the use of luteinizing hormone that releases hormone agonists. See Perlmutter M, Lepor H. Androgen deprivation therapy in the treatment of advanced prostate cancer Rev Urol. 2007; 9 (Suppl 1): S3-S8 and references therein.
Maximal androgen blockade is achieved by combining ADT with antiandrogen therapy. Antiandrogens compete with endogenous androgens, testosterone and dihydrotestosterone for binding to the ligand binding pocket of the androgen receptor (AR). AR belongs to the superfamily of nuclear hormone receptors and is mainly expressed in reproductive tissues and muscles. The binding of the ligand to the AR promotes its dissociation from the heat shock protein and other bands, resulting in the dimerization of the receptor, phosphorylation and subsequent displacement into the nucleus, where AR binds to the presence of prostate cells. An androgen responsive element of multiple gene regulatory regions that grow, survive, and differentiate.
The first non-steroidal antiandrogen flutamide was approved for PCa in 1989, and the structurally related compounds bicalutamide and nilutamide were used in 1995 and 1996, respectively. Launched in the year. Non-steroidal compounds are more advantageous than steroid anti-androgens in clinical applications because they do not interact with other steroid receptors and have improved oral bioavailability. In the structural classification of acetaminophen and androgen, bicalutamide is the most potent, most tolerant and the most important antiandrogen on the market. Bicalutamide is described in the patent literature, for example European Patent EP0100172. Certain arylamine derivatives have also been described as selective androgen receptor modulators in the documents WO 2008/011072 A2, WO 2010/116342 A2 and WO 2010/092546 A1.

Unfortunately, although ADT and antiandrogen treatment typically elicit an early favorable response, PCa then develops into a state in which androgen deprivation cannot control its malignancy despite the minimal amount of testosterone. This state is called castration resistant prostate cancer (CRPC) (or hormone resistant prostate cancer, HRPC) and is a lethal form of this disease. CRPC is believed to occur following genetic and/or epigenetic changes in prostate cancer cells and is characterized by reactivation of cancer cell growth in a hormone deprived environment that has adapted to the prostate.
The growth of cancer cells in CRPC is still dependent on the function of AR, and studies over the past decade have demonstrated that CRPC cells employ multiple mechanisms to reactivate AR. See Chen CD, Welsbie DS, Tran C, Baek SH, Chen R, Vessella R, Rosenfeld MG, Sawyers CL. Molecular determinants of resistance to antiandrogen therapy. Nat Med 2004 Jan; 10(1): 33-39 and references therein literature. The main mechanisms include amplification of the AR gene or upregulation of AR mRNA or protein, AR point mutations that allow AR to be activated by non-androgen ligands or even antiandrogens, co-activators of AR transcription and co-repressor expression Alteration, as well as AR selective splicing and sustained activation of variants. Therefore, drugs that standardize AR messages can still be effective in the prevention and treatment of CRPC.
The limited effect of currently available antiandrogen is most likely associated with incomplete AR inhibition in certain environments (Taplin ME. Drug insight: role of the androgen receptor in the development and progression of prostate cancer. Nat Clin Pract Oncol. 2007 Apr; 4(4): 236-244). Multiple molecular mechanisms can contribute to standard antiandrogen therapy. The use of an anti-androgen (e.g., bicalutamide) that targets the binding region of the AR ligand can result in the selection of prostate cancer cells with point mutations in the ligand binding region. In some cases these mutations can cause prostate cancer cells to convert antagonists into agonists. AR mutations were found in 10-40% of metastatic tumors. More than 70 mutations have been found in AR that result in increased basal activity or expanded ligand specificity of the receptor.
For example, mutations in the amino acid 877 from threonine to alanine are the mutations most commonly found in PCa patients, and in AR flutamide, cyprotenone (steroid-type androgen), progesterone and Estrogens are converted to agonistic effects. Mutation of amino acid 741 from tryptophan to leucine or cysteine causes bicalutamide to convert from antiandrogen to agonist (Hara T, Miyazaki J, Araki H, Yamaoka M, Kanzaki N , Kusaka M, Miyamoto M. Novel mutations of androgen receptor: a possible mechanism of bicalutamide withdrawal syndrome. Cancer Res. 2003 Jan 1; 63(1): 149-153).
In addition to AR point mutations, increased receptor levels can cause antiandrogen effects as agonists (Chen CD, Welsbie DS, Tran C, Baek SH, Chen R, Vessella R, Rosenfeld MG, Sawyers CL. Molecular determinants of resistance to Antiandrogen therapy. Nat Med 2004 Jan; 10(1): 33-39). The conversion of antagonist-agonists has significant clinical implications. Approximately 30% of men with progressive PCa experienced an abnormal decrease in serum prostate specific antigen levels after discontinuation of antiandrogen therapy.
It has been disappointing to date that the expected survival of CRPC treatment is estimated to be 7 to 16 months. Despite the recent addition of two new CRPC treatment options, the therapeutic prostate cancer vaccine, sipuleucel-T, and the new abiraterone acetate, a suppressor of the testosterone, still require a unique landmark for AR. Effective new pharmacy.
More specifically, there is a need for new anti-androgen compounds that are more effective than carotamide for combating endogenous androgenic activity on AR. There is also a need for new antiandrogen compounds that exhibit minimal agonism in AR. Importantly, there is a need for new antiandrogens that do not achieve pro-active activity in CRPC-associated mutant AR or in CRC-related treatments where AR is present in high amounts. In addition, there is a need for non-steroidal, non-toxic molecules with pharmacological properties useful for the treatment and prevention of BPH, PCa and CRPC.
It has now surprisingly been found that the arylamine derivatives according to the invention overcome one or more disadvantages associated with bicalutamide and other arylamine derivatives known in the art.

The present invention provides a novel arylamine derivative having the formula (I)

And stereoisomers thereof and pharmaceutically acceptable salts;


among them
R2 is halogen or trifluoromethyl;
R8 and R9 are selected from the group consisting of hydrogen, halogen and trifluoromethyl, and it is specified that at least one of R8 and R9 is other than hydrogen;
R11 is a branched C _3-5 -alkyl or cyclo-C _3-6 -alkyl group;
It is specified that when R11 is an isopentyl group, R2 is a trifluoromethyl group, and R9 is hydrogen, then R8 is not fluorine.
The invention is also related to pharmaceutical compositions comprising an effective amount of one or more of the arylamine derivatives of formula (I), or a pharmaceutically acceptable salt thereof, together with suitable carriers and conventional excipients.
Further, the present invention relates to the use of an arylamine derivative of the formula (I) or a pharmaceutically acceptable salt thereof as a medicament.
The invention also relates to the use of an arylamine derivative of the formula (I) or a pharmaceutically acceptable salt thereof for the treatment of androgen receptor-associated diseases.
Finally, the invention provides a procedure for the preparation of an arylamine derivative of formula (I).

The linaloamine of the formula (I) according to the invention has at least one asymmetric carbon atom, i.e., a carbon atom to which a hydroxyl group is bonded. Thus, the compound exists in a racemic form with an optically active form. All of these forms are included in the present invention.
Examples of branched C _3-5 -alkyl groups are isopropyl, iso- and tert-butyl and iso- and tert-amyl.
The term "cyclo-C _3-6 -alkyl" is used to mean cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.
The term "halogen" means fluorine, chlorine, bromine or iodine.
Preferred compounds of formula (I) are those wherein R2 is chloro or trifluoromethyl.
Further preferred compounds of formula (I) are those wherein R8 is chloro, fluoro or trifluoromethyl. Also preferred are those wherein R9 is trifluoromethyl, hydrogen or fluorine, preferably hydrogen or fluorine.
Further preferred compounds of formula (I) are those wherein one or both of R8 and R9 are independently selected from the group consisting of chlorine, fluorine and trifluoromethyl.
Preferred compounds of formula (I) are also those wherein R11 is tert-butyl, isopropyl, isopentyl, cyclopropyl, cyclopentyl or cyclohexyl, preferably isopropyl, isopentyl, Cyclopentyl or cyclohexyl.
Particularly preferred compounds of formula (I) are those wherein R2 is chloro, R8 is trifluoromethyl, R9 is hydrogen and R11 is cyclopentyl.
Examples of particularly preferred specific compounds are:
2-(4-Chlorophenyl)-N-[4-cyano-3-(trifluoromethyl)phenyl]-2-hydroxy-3-[(3-methylbutane)sulfonyl]propyl Guanamine
N-(3-chloro-4-cyanophenyl)-2-hydroxy-3-[(3-methylbutane)sulfonyl]-2-[4-(trifluoromethyl)phenyl]propanoid amine;
N-(3-chloro-4-cyanophenyl)-2-hydroxy-3-(propan-2-sulfonyl)-2-[4-(trifluoromethyl)phenyl]propanamine;
N-[4-Cyano-3-(trifluoromethyl)phenyl]-2-(4-fluorophenyl)-2-hydroxy-3-(propan-2-sulfonyl)propanamine;
N-(3-chloro-4-cyanophenyl)-3-(cyclohexylsulfonyl)-2-hydroxy-2-[4-(trifluoromethyl)phenyl]propanamine;
N-(3-chloro-4-cyanophenyl)-3-(cyclopentylsulfonyl)-2-hydroxy-2-[4-(trifluoromethyl)phenyl]propanamine;
2-(4-chlorophenyl)-N-[4-cyano-3-(trifluoromethyl)phenyl]-3-(cyclohexylsulfonyl)-2-hydroxypropionamide;
2-(4-chlorophenyl)-N-[4-cyano-3-(trifluoromethyl)phenyl]-3-(cyclopentylsulfonyl)-2-hydroxypropionamide;
2-(4-chlorophenyl)-N-[4-cyano-3-(trifluoromethyl)phenyl]-2-hydroxy-3-(propan-2-sulfonyl)propanamine;
N-(3-chloro-4-cyanophenyl)-2-(3,4-difluorophenyl)-2-hydroxy-3-[(3-methylbutane)sulfonyl]propanamine;
N-(3-chloro-4-cyanophenyl)-2-(3,4-difluorophenyl)-2-hydroxy-3-(propan-2-sulfonyl)propanamine;
N-(3-chloro-4-cyanophenyl)-2-(4-chlorophenyl)-2-hydroxy-3-[(3-methylbutane)sulfonyl]propanamine;
N-(3-chloro-4-cyanophenyl)-2-(4-chlorophenyl)-2-hydroxy-3-(propan-2-sulfonyl)propanamine;
N-[4-Cyano-3-(trifluoromethyl)phenyl]-2-hydroxy-3-[(3-methylbutane)sulfonyl]-2-[4-(trifluoromethyl) Phenyl]propanamine;
N-[4-Cyano-3-(trifluoromethyl)phenyl]-2-hydroxy-3-(propan-2-sulfonyl)-2-[4-(trifluoromethyl)phenyl] Propylamine;
N-[4-Cyano-3-(trifluoromethyl)phenyl]-2-(3,4-difluorophenyl)-2-hydroxy-3-[(3-methylbutane)sulfonate Propylamine
N-[4-Cyano-3-(trifluoromethyl)phenyl]-2-(3,4-difluorophenyl)-2-hydroxy-3-(propan-2-sulfonyl)propanoid amine;
N-(3-chloro-4-cyanophenyl)-2-(3,4-difluorophenyl)-2-hydroxy-3-(2-methylpropan-2-sulfonyl)propanamine;
2-(4-Chlorophenyl)-N-[4-cyano-3-(trifluoromethyl)phenyl]-2-hydroxy-3-(2-methylpropan-2-sulfonyl)propane Guanamine
N-(3-chloro-4-cyanophenyl)-3-(cyclopropanesulfonyl)-2-(3,4-difluorophenyl)-2-hydroxypropionamide;
2-(4-chlorophenyl)-N-[4-cyano-3-(trifluoromethyl)phenyl]-3-(cyclopropanesulfonyl)-2-hydroxypropionamide;
N-[4-Cyano-3-(trifluoromethyl)phenyl]-2-hydroxy-3-(2-methylpropan-2-sulfonyl)-2-[4-(trifluoromethyl) Phenyl]propanamine;
N-[4-Cyano-3-(trifluoromethyl)phenyl]-3-(cyclopropanesulfonyl)-2-hydroxy-2-[4-(trifluoromethyl)phenyl]propanoid amine;
N-[4-Cyano-3-(trifluoromethyl)phenyl]-2-(4-fluorophenyl)-2-hydroxy-3-(2-methylpropan-2-sulfonyl)propane Guanamine
N-[4-Cyano-3-(trifluoromethyl)phenyl]-3-(cyclopropanesulfonyl)-2-(4-fluorophenyl)-2-hydroxypropionamide;
N-[4-Cyano-3-(trifluoromethyl)phenyl]-2-(3,4-difluorophenyl)-2-hydroxy-3-(2-methylpropan-2-sulfonate) Acetylamine;
N-[4-Cyano-3-(trifluoromethyl)phenyl]-3-(cyclopropanesulfonyl)-2-(3,4-difluorophenyl)-2-hydroxypropionamide;
N-[4-Cyano-3-(trifluoromethyl)phenyl]-2-hydroxy-3-(2-methylpropan-2-sulfonyl)-2-[3-(trifluoromethyl) Phenyl]propanamine;
N-[4-Cyano-3-(trifluoromethyl)phenyl]-3-(cyclopropanesulfonyl)-2-hydroxy-2-[3-(trifluoromethyl)phenyl]propanoid amine;
N-(3-chloro-4-cyanophenyl)-2-hydroxy-3-(2-methylpropan-2-sulfonyl)-2-[3-(trifluoromethyl)phenyl]propanoid amine;
N-(3-chloro-4-cyanophenyl)-3-(cyclopropanesulfonyl)-2-hydroxy-2-[3-(trifluoromethyl)phenyl]propanamine;
N-(3-chloro-4-cyanophenyl)-2-(4-chlorophenyl)-2-hydroxy-3-(2-methylpropan-2-sulfonyl)propanamine;
N-(3-chloro-4-cyanophenyl)-2-(4-chlorophenyl)-3-(cyclopropanesulfonyl)-2-hydroxypropionamide;
N-[4-Cyano-3-(trifluoromethyl)phenyl]-2-hydroxy-3-(propan-2-sulfonyl)-2-[3-(trifluoromethyl)phenyl] Propylamine;
N-(3-chloro-4-cyanophenyl)-2-hydroxy-3-(propan-2-sulfonyl)-2-[3-(trifluoromethyl)phenyl]propanamine;
N-(3-chloro-4-cyanophenyl)-2-hydroxy-3-(2-methylpropan-2-sulfonyl)-2-[4-(trifluoromethyl)phenyl]propanoid amine;
N-(3-chloro-4-cyanophenyl)-3-(cyclopropanesulfonyl)-2-hydroxy-2-[4-(trifluoromethyl)phenyl]propanamine;
And pharmaceutically acceptable salts thereof.
Pharmaceutically acceptable salts and their preparation are well known in the art.
The linalylamine of the present invention can be prepared by the method described below. For example, a compound of formula (I) may be via an epoxy resin compound of formula (5) wherein R2, R8 and R9 are as defined above,

Prepared by reacting with a compound of formula (II),

Wherein R11 is as defined above, and the obtained sulfur-containing compound is oxidized to obtain a compound of the formula (I). This procedure is preferably carried out by the following reaction steps:

The general synthetic procedure uses the commercially available aniline, phenylacetic acid, mercaptans, phenols and amines as starting materials to synthesize the compounds of the invention.
General Synthetic Method for Intermediate (3) The corresponding phenylacetic acid (2) (3.89 mmol) was dissolved in dichloromethane and cooled to +5 - 0 °C in an ice bath. When the temperature was maintained at +5 - 0 ° C, 0.66 ml (2 equivalents) of grass sputum chloride was added dropwise to dichloromethane. After the completion of the addition, the ice bath was removed and the mixture was allowed to warm to room temperature (RT). After stirring for 4 hours, the mixture was cooled to 0 ° C and aniline (1) (3.89 mmol) was added in dimethylacetamide (10 ml). The resulting mixture was stirred at RT and monitored by TLC. After the reaction was completed, the mixture was poured into ice water and extracted with dichloromethane. The organic phase was washed with water and dried over Na ₂ SO ₄ and evaporated to afford (3).

General synthesis of intermediate (4) 1.7 mmol of (3), 0.075 g (1.8 equivalents) of paraformaldehyde with 0.412 g of K ₂ CO ₃ in NMP (N-methylpyrrolidone, 2 ml) Mixed in. The mixture was heated to 90 ° C and stirred for 3 hours. After cooling to RT, 10 ml of water was added and the mixture was extracted with diisopropyl ether (2 x 10 ml). The organic phase was washed with water (1 x 10 ml) and evaporated to provide (4). The product was used in the synthesis of (5) without further purification.
General Synthetic Method of Intermediate (5) 1.0 mmol of the intermediate product (4) and 10 mg of 2,6-di-tert-butyl-4-methylphenol were dissolved in CH ₂ Cl ₂ (20 ml). Add 0.5 g (2 equivalents) of MCPBA. The mixture was stirred overnight at RT. The mixture was extracted with Na ₂ CO ₂ and water. The organic phase is evaporated under vacuum to provide the epoxide (5). The product was used in the synthesis of (6) without further purification.
General Synthetic Method of (6) 2.2 mmol (1.5 equivalents) of corresponding thiophenol or phenol is added to THF (7.5 ml) in 3.0 (2 equivalents) in THF (5 ml) at 0 °C. Methyl K ₂ CO ₃ . The mixture was stirred at 0 ° C for 30 min. 1.5 mmol of epoxide (5) in THF (7.5 ml) was added at 0 °C. The resulting mixture was stirred at RT for 2 h, heated to 50 ° C and stirred for 12 h. After the reaction was cooled, the reaction was quenched with water. The resulting mixture was extracted with AcOEt. The organic phase was concentrated to give a crude material which was used for the synthesis of (7) without further purification. In the case where the phenol is used in the reaction, the product is purified using flash chromatography.
General Synthetic Method of (7) 0.45 mmol of (6) was dissolved in CH ₂ Cl ₂ (20 ml). MCPBA (0.90 mmol, 2 equivalents) was added and the mixture was stirred at RT. After completion of the reaction under TLC monitoring, the reaction was quenched with saturated sodium sulfite solution in water and extracted with dichloromethane. The organic layer was washed with a saturated aqueous solution of sodium sulphate, dried over Na ₂ SO ₄ and evaporated. The product was purified using flash chromatography.
Preparation of Cyclopropyl Mercaptan Cyclopropyl thiol was prepared according to the method described in JACS 1992, 114(9), 3492-3499.
EXAMPLES The compounds listed in Table 1 below were prepared using the synthetic procedures described above and illustrate the invention.







General Description of Pharmacological Properties of the Compounds of the Invention The linaloamine derivatives of the present invention exhibit high antagonistic activity at AR. Antagonistic activity at AR means the strength of the compound to compete and/or inhibit the activity of natural AR ligands such as dihydrosterolone (DHT) and testosterone. The present invention provides a compound having antagonistic activity in AR to compete for and/or inhibit the activity of a non-natural AR ligand which is, for example, a synthetic androgen or antibiotic used as a drug (but which may cause harmful side effects) Androgen.
Further, the present invention provides compounds which exhibit potent anti-androgenic activity in a dose-dependent manner. The main disadvantage of bicalutamide is incomplete AR antagonism. In the case of bicalutamide, the increased concentration does not provide significant additional benefits. It may be desirable to use anti-androgens that are more potent than bicalutamide to treat advanced PCa characterized by elevated AR levels, and therefore there is a need for an effective anti-androgen that can compensate for elevated AR levels in a dose-dependent manner. The present invention provides a compound that exerts minimal stimulatory effects in AR.
The compounds of the invention may be used to treat AR related diseases, such as BPH and PCa. This compound can also be used to treat CRPC. Further, the compound can be used in combination with other antiandrogen therapies.
The compounds of the invention do not achieve pro-active activity in CRPC-associated mutations. All mutations affecting the development, evolution or severity of the disease are addressed by CRPC-associated mutations. CRPC-associated mutations may have been attributed to the potentiation of androgen deprivation induction by prostate cancer cells bearing the mutation. For example, mention is made of mutations of tryptophan 741 to leucine or to cysteine and mutations of sulphite 877 to alanine.
When the amount of AR is increased, the compound of the present invention retains its antagonistic activity.
The following tests and results are provided to demonstrate the invention in an illustrative manner and are not to be construed as limiting the scope of the invention. Further, the concentration of the compound in the test is exemplary and should not be taken as a limitation. Those skilled in the art can define pharmaceutically relevant concentrations by methods known in the art.
Experiments to demonstrate the efficacy of the compounds of the invention as anti-androgens and demonstrate that the compounds of the invention are efficacious in first-line antiandrogens (eg, flutamide or bicalutamide, BIC) that are known to be licensed for clinical use. A series of in vitro studies were designed to retain its antagonistic activity under active conditions. These studies measure AR transactivation based on the use of reporter gene assays, which are well established gold standard tests in AR studies. Depending on the presence or absence of a natural AR ligand (eg, testosterone), this reporter gene assay can be used to determine both the antagonistic and agonistic activities of the compound. BIC was used as a reference compound in all studies, representing the currently available standard antiandrogen therapy.
AR transactivation assay COS-1 cells (American Type Culture Collection, ATCC) were cultured in Dulbecco's supplemented with 10% fetal bovine serum (FBS), penicillin (6.25 U/ml) and streptomycin (6.25 μg/ml). Modified Eagle Medium (DMEM) and inoculated into a 48-well plate (50,000 cells/well) one day prior to transfection. The transfected broth containing 2.5% charcoal-adsorbed FBS in DMEM was replaced 4 h before transfection. The plastids (pPB-286/+32-LUC; PB, probasin promoter) and 5 ng AR were expressed in 50 ng of cold light enzyme (LUC) using TransIT-LT1 reagent (Mirus Bio Corporation) according to the manufacturer's instructions. (pSG5-hAR) and 5 ng of pCMVβ (control group for an internal β-galactosidase for transfection efficiency and cell growth) transfected cells. After one day of transfection, the three replicate wells received (i) vehicle (EtOH-DMSO), (ii) 50 nM guanosterone (reference agonist from Makor or Steraloids Inc.), (iii) increased concentration of BIC ( Reference antagonist) or (iv) only compounds of the invention (to test for agonism) or (v) increased concentrations of BIC (reference antagonist) or (vi) in a competitive environment (50 nM; to test for testosterone induction The AR transcriptional antagonism of the compounds of the invention and a reference agonist. After 18 h, reporter gene activity (LUC and β-galactosidase) was determined according to standard methods. Data are expressed as the relative LUC activity of the given compound relative to the reference test item activity (=100%) (cold light enzyme light unit divided by β-galactosidase A420 _nm versus control group as transfection efficiency).
Alternatively, a commercial human AR reporter test system (INDIGO Biosciences) was used. In this assay, non-human mammalian cells were designed to express human WT AR along with the LUC reporter gene linked to the AR response promoter. 400 pM 6-α-FI testosterone and FIT were used as reference agonists in a competitive environment. Two reported gene systems lead to comparable data.
The agonistic effect in WT AR measures the WT AR agonism of the compounds of the invention by the AR transactivation assay in COS-1 cells by transfecting the cells as described above with exposure to the test compound alone. Use testosterone as a reference agonist. The relative LUC activity representing the degree of AR activation was measured. The reaction obtained from the reference agonist was set to 100%. The compounds of the invention do not show an agonistic effect in WT AR.
Antagonism in wild-type (WT) AR The antagonism of WT AR of the compounds of the present invention was measured in an AR transactivation assay of COS-1 cells in a competitive environment as described above using sterolone as a reference agonist. The INDIGO Bioscience's Human AR reporter test system was alternatively utilized. A known antiandrogen BIC was used as a reference antagonist. The relative LUC activity representing AR-dependent transcription obtained by exposure to only the reference agonist was set to 100%. The compounds of the invention are potent antagonists in WT AR (Table 2).


One of the major limitations in the use of currently available antiandrogens such as flutamide and BIC is the antagonist-agonist transition observed in mutant AR.
The agonistic effect of the W741L mutant AR In addition to the use of the AR expression vector with the W741L mutation in place of the WT AR, the agonist effect of the W741L AR of the compound of the present invention was measured as described above in the AR transactivation assay of COS-1 cells. The transfected cells are individually exposed to the test compound. BIC was used as a reference compound. As reported in the literature, BIC acts as an agonist in this mutant AR variant and sets the BCI-induced relative LUC activity representing AR-dependent transcription to 100%. The compounds of the invention showed no agonistic effects in W741L AR (Table 3).
The agonistic effect of T877A mutant AR In addition to the use of the AR expression vector with the T877A mutation, the agonist effect of the T877A AR of the compound of the present invention was measured as described above in the AR transactivation assay of COS-1 cells. The transfected cells are individually exposed to the test compound. Clottenone was used as a reference agonist, and its relative LUC activity representing AR-dependent transcription was set to 100%. The compounds of the invention showed no agonistic effects in T877A AR (Table 3).

Gene Expression in VCaP Cells The ability of the compounds of the invention to inhibit the expression of the AR marker gene was investigated using quantitative RT-PCR. VCaP cells (ATCC) were seeded into 12-well plates (3 x 10 ⁵ cells/well) and either (i) vehicle (EtOH-DMSO), or (ii) 1 nM R1881 (reference agonist, Perkin-Elmer) Or (iii) increasing concentrations of BIC (reference antagonist), or (iv) test compound and reference agonist (1 nM) (all final concentrations) to treat three replicate wells. After 18 h, total RNA was extracted using TRIzolR reagent (Invitrogen Life Technologies) and converted to cDNA using the Transcriptor First Strand cDNA synthesis kit (Roche Diagnostics GmbH) according to the manufacturer's instructions. cDNA was used as a template for RT-qPCR using Mx3000P Real-Time PCR System (Stratagene), FastStart SYBR Green Master Mix (Roche), and specific primers for the AR marker gene, PSA, TMPRSS2 and FKBP51. The amount of GAPDH mRNA analyzed was used to normalize the amount of total RNA between samples. Using the formula ^{2 - (ΔΔ Ct)} fold change is calculated (ligand-induced), which is ΔΔCt ACt _{(ligand) -ΔCt (EtOH-DMSO),} ΔCt is Ct _{(Gene X)} -Ct _(GAPDH), and Ct is the threshold crossings The loop. Gene expression data is expressed as the relative amount of mRNA of each gene of the given compound (the amount of mRNA of the gene of interest divided by the amount of mRNA of GAPDH). The compounds of the invention efficiently silence the AR marker gene in VCaP cells.
LNCaP Proliferation Assay The ability of the compounds of the invention to inhibit the growth of prostate cancer cells was investigated in the androgen sensitive human prostate adenocarcinoma cell line LNCaP (ATCC). It is also possible to genetically modify LNCaP cells to overexpress AR, thus mimicking CRPC. The cells were seeded in 96-well plates (5000 cells/well) and cultured for 24 h. Taking (i) vehicle (DMSO), or (ii) 0.1 nM R1881 (reference agonist, Perkin-Elmer) or (iii) increasing concentration of BIC (reference antagonist), or (iv) test compound together with reference The efflux (0.1 nM) (all final concentrations) was treated for six replicate wells for 5 days. Promega's Cell Titer 96 ^R AQ _ueous One Solution Cell Proliferation Assay kits were used on Days 0, 1, 3, and 5 to measure LNCaP cell proliferation according to the manufacturer's instructions. 20 μl of Cell Titer reagent was added to 100 μl of cell culture medium per well, and the cells were allowed to grow in an incubator for one hour. The medium was transferred to the wells of the measuring disk and the absorbance at 492 nm was recorded. The compounds of the invention inhibit LNCaP proliferation.
The compounds of the invention exhibit little or no agonistic activity on androgen receptors. Because these compounds are potent AR antagonists, they are not only useful for the treatment of prostate cancer, but also for other androgen receptor-related conditions and diseases such as benign prostatic hyperplasia, hair loss, acne, hirsutism, and male hypersexuality. Or polycystic ovarian syndrome.
The compounds of the invention may be used alone or in combination, i.e., simultaneously, separately or sequentially in combination with other active agents.
Since it relates to the treatment of cancer, the compounds of the invention are optimally used alone or in combination with anti-androgen cancer treatment. Such compounds may also be combined with agents that inhibit the production of circulating steroids, such as LHRH agonists or antagonists, or with surgical castration.
The invention also contemplates the use of an anti-estrogen and/or aromatase inhibitor in combination with a compound of the invention, for example, to help alleviate side effects associated with anti-androgen therapy, such as gynecomastia ).
AR belongs to the superfamily of nuclear receptors, and can also be used as a drug design of other nuclear hormone receptors (such as estrogen receptor or peroxisome proliferator-activated receptor). Bracket. Therefore, the compounds of the present invention can be further optimized for the treatment of other symptoms and diseases, such as ovarian cancer, breast cancer, diabetes, heart disease, peripheral and central nervous system in which nuclear receptors play a role. Metabolic related diseases.
The compounds of the invention may be administered by intravenous injection, via injection to tissue, intraperitoneal injection, orally or by nasal administration. The composition may have a form selected from the group consisting of a solution, a dispersion, a suspension, a powder, a capsule, a tablet, a pill, a controlled release capsule, a controlled release tablet, and a controlled release pellet.

Claims (15)

An arylamine derivative having the formula (I)

And stereoisomers thereof and pharmaceutically acceptable salts;
among them
R2 is halogen or trifluoromethyl;
R8 and R9 are selected from the group consisting of hydrogen, halogen and trifluoromethyl, and it is stated that at least one of R8 and R9 is not hydrogen;
R11 is a branched C _3-5 -alkyl or cyclo-C _3-6 -alkyl group;
It is specified that when R11 is an isopentyl group, R2 is a trifluoromethyl group, and R9 is hydrogen, then R8 is not fluorine. An arylamine derivative as described in claim 1 wherein R2 is chloro or trifluoromethyl. An arylamine derivative as described in claim 1 or 2, wherein R8 is chloro, fluoro or trifluoromethyl. The arylamine derivative according to any one of claims 1 to 3, wherein R9 is trifluoromethyl, hydrogen or fluorine. An arylamine derivative as described in claim 1, wherein R11 is tert-butyl, isopropyl, isopentyl, cyclopropyl, cyclopentyl or cyclohexyl. An arylamine derivative as described in claim 1, wherein the linoleamide derivative is selected from the group consisting of:
2-(4-Chlorophenyl)-N-[4-cyano-3-(trifluoromethyl)phenyl]-2-hydroxy-3-[(3-methylbutane)sulfonyl]propyl Guanamine
N-(3-chloro-4-cyanophenyl)-2-hydroxy-3-[(3-methylbutane)sulfonyl]-2-[4-(trifluoromethyl)phenyl]propanoid amine;
N-(3-chloro-4-cyanophenyl)-2-hydroxy-3-(propan-2-sulfonyl)-2-[4-(trifluoromethyl)phenyl]propanamine;
N-[4-Cyano-3-(trifluoromethyl)phenyl]-2-(4-fluorophenyl)-2-hydroxy-3-(propan-2-sulfonyl)propanamine;
N-(3-chloro-4-cyanophenyl)-3-(cyclohexylsulfonyl)-2-hydroxy-2-[4-(trifluoromethyl)phenyl]propanamine;
N-(3-chloro-4-cyanophenyl)-3-(cyclopentylsulfonyl)-2-hydroxy-2-[4-(trifluoromethyl)phenyl]propanamine;
2-(4-chlorophenyl)-N-[4-cyano-3-(trifluoromethyl)phenyl]-3-(cyclohexylsulfonyl)-2-hydroxypropionamide;
2-(4-chlorophenyl)-N-[4-cyano-3-(trifluoromethyl)phenyl]-3-(cyclopentylsulfonyl)-2-hydroxypropionamide;
2-(4-chlorophenyl)-N-[4-cyano-3-(trifluoromethyl)phenyl]-2-hydroxy-3-(propan-2-sulfonyl)propanamine;
N-(3-chloro-4-cyanophenyl)-2-(3,4-difluorophenyl)-2-hydroxy-3-[(3-methylbutane)sulfonyl]propanamine;
N-(3-chloro-4-cyanophenyl)-2-(3,4-difluorophenyl)-2-hydroxy-3-(propan-2-sulfonyl)propanamine;
N-(3-chloro-4-cyanophenyl)-2-(4-chlorophenyl)-2-hydroxy-3-[(3-methylbutane)sulfonyl]propanamine;
N-(3-chloro-4-cyanophenyl)-2-(4-chlorophenyl)-2-hydroxy-3-(propan-2-sulfonyl)propanamine;
N-[4-Cyano-3-(trifluoromethyl)phenyl]-2-hydroxy-3-[(3-methylbutane)-sulfonyl]-2-[4-(trifluoromethyl) Phenyl)propanamine;
N-[4-Cyano-3-(trifluoromethyl)phenyl]-2-hydroxy-3-(propan-2-sulfonyl)-2-[4-(trifluoromethyl)phenyl] Propylamine;
N-[4-Cyano-3-(trifluoromethyl)phenyl]-2-(3,4-difluorophenyl)-2-hydroxy-3-[(3-methylbutane)sulfonate Propylamine
N-[4-Cyano-3-(trifluoromethyl)phenyl]-2-(3,4-difluorophenyl)-2-hydroxy-3-(propan-2-sulfonyl)propanoid amine;
N-(3-chloro-4-cyanophenyl)-2-(3,4-difluorophenyl)-2-hydroxy-3-(2-methylpropan-2-sulfonyl)propanamine;
2-(4-Chlorophenyl)-N-[4-cyano-3-(trifluoromethyl)phenyl]-2-hydroxy-3-(2-methylpropan-2-sulfonyl)propane Guanamine
N-(3-chloro-4-cyanophenyl)-3-(cyclopropanesulfonyl)-2-(3,4-difluorophenyl)-2-hydroxypropionamide;
2-(4-chlorophenyl)-N-[4-cyano-3-(trifluoromethyl)phenyl]-3-(cyclopropanesulfonyl)-2-hydroxypropionamide;
N-[4-Cyano-3-(trifluoromethyl)phenyl]-2-hydroxy-3-(2-methylpropan-2-sulfonyl)-2-[4-(trifluoromethyl) Phenyl]propanamine;
N-[4-Cyano-3-(trifluoromethyl)phenyl]-3-(cyclopropanesulfonyl)-2-hydroxy-2-[4-(trifluoromethyl)phenyl]propanoid amine;
N-[4-Cyano-3-(trifluoromethyl)phenyl]-2-(4-fluorophenyl)-2-hydroxy-3-(2-methylpropan-2-sulfonyl)propane Guanamine
N-[4-Cyano-3-(trifluoromethyl)phenyl]-3-(cyclopropanesulfonyl)-2-(4-fluorophenyl)-2-hydroxypropionamide;
N-[4-Cyano-3-(trifluoromethyl)phenyl]-2-(3,4-difluorophenyl)-2-hydroxy-3-(2-methylpropan-2-sulfonate) Acetylamine;
N-[4-Cyano-3-(trifluoromethyl)phenyl]-3-(cyclopropanesulfonyl)-2-(3,4-difluorophenyl)-2-hydroxypropionamide;
N-[4-Cyano-3-(trifluoromethyl)phenyl]-2-hydroxy-3-(2-methylpropan-2-sulfonyl)-2-[3-(trifluoromethyl) Phenyl]propanamine;
N-[4-Cyano-3-(trifluoromethyl)phenyl]-3-(cyclopropanesulfonyl)-2-hydroxy-2-[3-(trifluoromethyl)phenyl]propanoid amine;
N-(3-chloro-4-cyanophenyl)-2-hydroxy-3-(2-methylpropan-2-sulfonyl)-2-[3-(trifluoromethyl)phenyl]propanoid amine;
N-(3-chloro-4-cyanophenyl)-3-(cyclopropanesulfonyl)-2-hydroxy-2-[3-(trifluoromethyl)phenyl]propanamine;
N-(3-chloro-4-cyanophenyl)-2-(4-chlorophenyl)-2-hydroxy-3-(2-methylpropan-2-sulfonyl)propanamine;
N-(3-chloro-4-cyanophenyl)-2-(4-chlorophenyl)-3-(cyclopropanesulfonyl)-2-hydroxypropionamide;
N-[4-Cyano-3-(trifluoromethyl)phenyl]-2-hydroxy-3-(propan-2-sulfonyl)-2-[3-(trifluoromethyl)phenyl] Propylamine;
N-(3-chloro-4-cyanophenyl)-2-hydroxy-3-(propan-2-sulfonyl)-2-[3-(trifluoromethyl)phenyl]propanamine;
N-(3-chloro-4-cyanophenyl)-2-hydroxy-3-(2-methylpropan-2-sulfonyl)-2-[4-(trifluoromethyl)phenyl]propanoid amine;
N-(3-chloro-4-cyanophenyl)-3-(cyclopropanesulfonyl)-2-hydroxy-2-[4-(trifluoromethyl)phenyl]propanamine;
And pharmaceutically acceptable salts thereof. A pharmaceutical composition comprising an effective amount of one or more linalylamine derivatives or a pharmaceutically acceptable salt thereof according to any one of claims 1-6, and a suitable one Carrier with traditional excipients. The use of the arylamine derivative or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 6 as a medicament. Use of an arylamine derivative or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 6 for the treatment of androgen receptor-related disorders. An arylamine derivative or a pharmaceutically acceptable salt thereof for use according to the scope of claim 9 wherein the disease is benign prostatic hyperplasia. An arylamine derivative or a pharmaceutically acceptable salt thereof for use as claimed in claim 9 wherein the disorder is cancer. An arylamine derivative or a pharmaceutically acceptable salt thereof for use according to the scope of claim 11 wherein the cancer is selected from the group consisting of prostate cancer and castration-resistant prostate cancer Group of. An arylamine derivative or a pharmaceutically acceptable salt thereof for use according to any one of claims 9 to 12, wherein the compound is simultaneously, separately or continuously with another active agent Administered. A process for preparing an arylamine derivative of the formula (I) as defined in the first aspect of the patent application, which comprises an epoxy resin compound of the formula (5) wherein R2, R8 and R9 are as Whoever is defined in the first paragraph of the patent application,

Reacting with a compound of formula (II)

Wherein R11 is as defined in claim 1 of the patent application, and the obtained sulfur-containing compound is oxidized to obtain a compound of the formula (I). The procedure as described in claim 14, wherein the program is carried out by the following reaction steps